猪Gasp1和Gasp2基因对C2C12细胞分化的影响

doi:10.15933/j.cnki.1004-3268.2018.03.024

河南农业科学 ›› 2018, Vol. 47 ›› Issue (3): 121-127.DOI: 10.15933/j.cnki.1004-3268.2018.03.024

猪Gasp1和Gasp2基因对C2C12细胞分化的影响

樊爽爽，张雪梅，刘晓贺，刘姣扬，明胜利，褚贝贝，杨国宇，王江

河南农业大学农业部动物生化与营养重点开放实验室，河南郑州 450002

收稿日期:2017-08-16 出版日期:2018-03-15 发布日期:2018-03-15
通讯作者: 王江(1982-)，男，陕西蒲城人，讲师，博士，主要从事动物生物化学研究工作。E-mail：nowingdream@126.com
作者简介:樊爽爽(1991-)，女，河南荥阳人，在读硕士研究生，研究方向：动物生物化学。E-mail：990461231@qq.com
基金资助:
农业部转基因重大专项（2014ZX0801015B）；农业部948重点计划(2011-G35)

Received:2017-08-16 Published:2018-03-15 Online:2018-03-15

摘要/Abstract

摘要： 为研究猪Gasp1、Gasp2对成肌细胞分化的影响，采用RT-PCR技术从猪肌肉组织中扩增Gasp1和Gasp2基因片段，将其克隆入phiC31载体，构建重组质粒 phiC31-Gasp1和phiC31-Gasp2。将重组质粒转染至C2C12小鼠成肌细胞，经嘌呤霉素筛选在荧光显微镜下观察，获得稳定表达Gasp1和Gasp2基因的细胞系后，应用CCK-8法检测细胞增殖能力。分别在细胞分化1、2、3、4、5 d时收集细胞，采用实时荧光定量PCR检测MyoD、MyoG、P21及MHC肌细胞分化相关基因的表达变化，进而分析Gasp1和Gasp2基因对肌肉分化的影响。结果显示，本试验成功克隆了猪Gasp1和Gasp2基因，构建了phiC31- Gasp1和phiC31-Gasp2重组质粒，荧光显微镜下可见phiC31载体和Gasp1、Gasp2基因共转染的C2C12细胞表达强烈的绿色荧光，且对细胞的生长增殖无明显影响；实时荧光定量PCR检测结果表明，猪Gasp1和Gasp2可以促进细胞分化因子MyoD、MyoG、P21、MHC基因的表达。以上结果表明，Gasp1、Gasp2基因可以促进肌肉分化。

关键词: 猪Gasp1和Gasp2, 基因克隆, 真核表达, C2C12细胞分化

中图分类号:

S828

樊爽爽，张雪梅，刘晓贺，刘姣扬，明胜利，褚贝贝，杨国宇，王江. 猪Gasp1和Gasp2基因对C2C12细胞分化的影响[J]. 河南农业科学, 2018, 47(3): 121-127.

参考文献

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猪Gasp1和Gasp2基因对C2C12细胞分化的影响

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